Heterocyclic compounds as RSV inhibitors

ABSTRACT

The present invention discloses compounds of Formula (I), or pharmaceutically acceptable salts, esters, or prodrugs thereof:which inhibit Respiratory Syncytial Virus (RSV). The present invention further relates to pharmaceutical compositions comprising the aforementioned compounds for administration to a subject suffering from RSV infection. The invention also relates to methods of treating an RSV infection in a subject by administering a pharmaceutical composition comprising the compounds of the present invention.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No.62/831,470, filed on Apr. 9, 2019. The entire teachings of the aboveapplication are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates generally to compounds and pharmaceuticalcompositions useful as Respiratory Syncytial Virus (RSV) inhibitors.

BACKGROUND OF THE INVENTION

Human respiratory syncytial virus (HRSV) is a negative-sense, singlestranded, RNA paramyxovirus (KM. Empey, et al., Rev. Anti-InfectiveAgents, 2010, 50(1 May), 1258-1267). RSV is the leading cause of acutelower respiratory tract infections (ALRI) and affects patients of allages. The symptoms in adults are usually not severe and are typicallyanalogous to a mild cold. However, in infants and toddlers the virus cancause lower respiratory tract infections including bronchiolitis orpneumonia with many of them requiring hospitalization. Nearly allchildren have been infected by age 3. There are known high-risk groupsthat infection with RSV is more likely to progress into the ALRI.Premature infants and/or infants suffering from lung or cardiac diseaseare at the highest risk to develop ALRI. Additional high-risk groupsinclude the elderly, adults with chronic heart and/or lung disease, stemcell transplant patients and the immunosuppressed.

Currently, there is no vaccine available to prevent HRSV infection.Palivizumab is a monoclonal antibody that is used prophylactically toprevent HRSV infection in high risk infants, e.g. premature infants, andinfants with cardiac and/or lung disease. The high cost of palivizumabtreatment limits its use for general purposes. Ribavirin has also beenused to treat HRSV infections but its effectiveness is limited. There isa major medical need for new and effective HRSV treatments that can beused generally by all population types and ages.

There have been several RSV fusion inhibitors that have been disclosedin the following publications: WO2010/103306, WO2012/068622,WO2013/096681, WO2014/060411, WO2013/186995, WO2013/186334, WO2013/186332, WO 2012 080451, WO 2012/080450, WO2012/080449, WO2012/080447, WO 2012/080446, WO 2016/055792, WO 2016/097761, and J. Med.Chem. 2015, 58, 1630-1643. Examples of other N-protein inhibitors fortreatment of HRSV have been disclosed in the following publications: WO2004/026843, J. Med Chem. 2006, 49, 2311-2319, and J. Med Chem. 2007,50, 1685-1692. Examples of L-protein inhibitors for HRSV have beendisclosed in the following publications: WO 2011/005842, WO 2005/042530,Antiviral Res. 2005, 65, 125-131, and Bioorg. Med Chem. Lett. 2013, 23,6789-6793, Bioorg. Med Chem. Lett. 2017, 27, 2201-2206. Examples ofnucleosides/polymerase inhibitors have been disclosed in the followingpublications: WO 2013/242525 and J. Med Chem. 2015, 58, 1862-1878.

There is a need for the development of effective treatments for HRSV.The present invention has identified these novel compounds and theirinhibitory activity against HRSV. The invention includes methods toprepare the compounds as well as methods of using these compounds totreat disease.

SUMMARY OF THE INVENTION

The present invention provides compounds represented by Formula (I), andpharmaceutically acceptable salts, esters and prodrugs thereof that canbe used to treat or prevent viral (particularly HRSV) infection:

wherein:A is selected from the group consisting of:

1) optionally substituted aryl;

2) optionally substituted heteroaryl;

3) optionally substituted —C₃-C₁₂ cycloalkyl;

4) optionally substituted —C₃-C₁₂ cycloalkenyl; and

5) optionally substituted 3- to 12-membered heterocycloalkyl;

B is selected from the group consisting of:

1) optionally substituted aryl;

2) optionally substituted heteroaryl;

3) optionally substituted —C₃-C₁₂ cycloalkyl;

4) optionally substituted —C₃-C₁₂ cycloalkenyl; and

5) optionally substituted 3- to 12-membered heterocycloalkyl;

L is absent, —CONH—, —NHCO—, —NHCO₂—, or —NHS(O)₂—; preferably L is—CONH—;

Each R₁, R₂, and R₃ is independently selected from the group consistingof:

1) halogen;

2) cyano;

3) nitro;

4) optionally substituted —C₁-C₈ alkoxy;

5) optionally substituted —C₁-C₈ alkyl;

6) optionally substituted —C₂-C₈ alkenyl;

7) optionally substituted —C₂-C₈ alkynyl;

8) optionally substituted —C₃-C₆ cycloalkyl;

9) optionally substituted —C₃-C₆ cycloalkenyl; and

10) optionally substituted 3- to 6-membered heterocycloalkyl;

R₄ is selected from the group consisting of:

1) hydrogen;

2) halogen;

3) cyano;

4) nitro;

5) optionally substituted —C₁-C₈ alkoxy;

6) optionally substituted —C₁-C₈ alkyl;

7) optionally substituted —C₂-C₈ alkenyl;

8) optionally substituted —C₂-C₈ alkynyl;

9) optionally substituted —C₃-C₈ cycloalkyl;

10) optionally substituted —C₃-C₈ cycloalkenyl; and

11) optionally substituted 3- to 8-membered heterocycloalkyl;

n is 0, 1, 2, 3 or 4;

m is 0, 1, 2, 3 or 4; and

v is 0, 1, 2, or 3.

Each preferred group stated above can be taken in combination with one,any or all other preferred groups.

DETAILED DESCRIPTION OF THE INVENTION

In one embodiment the present invention provides a compound representedby Formula (I) as described above, or a pharmaceutically acceptablesalt, ester or prodrug thereof.

In certain embodiments of the compounds of Formula (I), n is 1 to 4 andeach R₁ is independently halogen, optionally substituted —C₁-C₃ alkoxy,or optionally substituted —C₁-C₃ alkyl.

In certain embodiments of the compounds of Formula (I), n is 1 or 2 andeach R₁ is independently F, optionally substituted —OCH₃, or optionallysubstituted —CH₃.

In certain embodiments of the compounds of Formula (I), n is 0.

In certain embodiments of the compounds of Formula (I), m is 1 to 4 andeach R₂ is independently halogen, optionally substituted —C₁-C₃ alkoxy,or optionally substituted —C₁-C₃ alkyl.

In certain embodiments of the compounds of Formula (I), m is 1 or 2 andeach R₂ is independently F, optionally substituted —OCH₃, or optionallysubstituted —CH₃.

In certain embodiments of the compounds of Formula (I), m is 0.

In certain embodiments of the compounds of Formula (I), wherein n is 1,m is 0 and R₁ is F.

In certain embodiments of the compounds of Formula (I), v is 1 to 3, andeach R₃ is independently halogen, optionally substituted —C₁-C₃ alkoxy,optionally substituted —C₁-C₃ alkyl, or optionally substituted —C₃-C₆cycloalkyl.

In certain embodiments of the compounds of Formula (I), v is 1 and R₃ isF, optionally substituted —OCH₃, optionally substituted —CH₃, oroptionally substituted cyclopropyl.

In certain embodiments of the compounds of Formula (I), v is 0.

In certain embodiments of the compounds of Formula (I), n is 0 and m is0.

In certain embodiments of the compounds of Formula (I), n is 0, m is 0,and v is 0.

In certain embodiments of the compounds of Formula (I), R₄ is halogen,optionally substituted —C₁-C₃ alkoxy, or optionally substituted —C₁-C₃alkyl.

In certain embodiments of the compounds of Formula (I), R₄ is H, F,optionally substituted —OCH₃, or optionally substituted —CH₃. In certainembodiments, R₄ is —CH₃, —CF₃ or —CHF₂.

In certain embodiments of the compounds of Formula (I), L is absent,—CONH—, or —NHCO—. In certain embodiments, L is —CONH—.

In certain embodiments of the compounds of Formula (I), A is optionallysubstituted aryl; preferably A is optionally substituted phenyl. Theoptional substituents are preferably independently selected from, butnot limited to, halogen, —CN, —OH, —NH₂, —NO₂, —CH₃, —CF₃, —OCH₃, —OCF₃,—SO₂CH₃, —CH₂N(CH₃)₂, and —C(O)CH₃. In preferred embodiments, there are0 to 2 substituents and, more preferably, 0 or 1 substituent.

In certain embodiments of the compounds of Formula (I), A is optionallysubstituted heteroaryl; preferably A is optionally substitutedthiophenyl. When present the substituent or substituents are preferablyindependently selected from halogen, —CN, —OH, —NH₂, —NO₂, —CH₃, CF₃,—OCH₃, —OCF₃, —SO₂CH₃, —CH₂N(CH₃)₂, and —C(O)CH₃. In preferredembodiments, there are 0 to 2 substituents and, more preferably, 0 or 1substituent.

In certain embodiments of the compounds of Formula (I), B is optionallysubstituted aryl; preferably B is optionally substituted phenyl. Theoptional substituents are preferably independently selected from, butnot limited to, halogen, —CN, —OH, —NH₂, —NO₂, —CH₃, —CF₃, —OCH₃, —OCF₃,—SO₂CH₃, —CH₂N(CH₃)₂, and —C(O)CH₃. In preferred embodiments, there are0 to 2 substituents and, more preferably, 0 or 1 substituent.

In certain embodiments of the compounds of Formula (I), B is optionallysubstituted heteroaryl. When present the substituent or substituents arepreferably independently selected from halogen, —CN, —OH, —NH₂, —NO₂,—CH₃, —CF₃, —OCH₃, —OCF₃, —SO₂CH₃, —CH₂N(CH₃)₂, and —C(O)CH₃. Inpreferred embodiments, there are 0 to 2 substituents and, morepreferably, 0 or 1 substituent.

In certain embodiments of the compounds of Formula (I), B is optionallysubstituted —C₃-C₁₂ cycloalkyl. When present the substituent orsubstituents are preferably independently selected from halogen, —CN,—OH, —NH₂, —NO₂, —CH₃, —CF₃, —OCH₃, —OCF₃, —SO₂CH₃, —CH₂N(CH₃)₂, and—C(O)CH₃. In preferred embodiments, there are 0 to 2 substituents and,more preferably, 0 or 1 substituent.

In another embodiment, the invention provides a compound represented byFormula (IIa)˜(IIc) or a pharmaceutically acceptable salt, ester orprodrug thereof:

wherein A, B, R₁, R₂, R₃, R₄, n, m, and v are as previously defined.

In another embodiment, the invention provides a compound represented byone of Formulas (IIIa)˜(IIIc) or a pharmaceutically acceptable salt,ester or prodrug thereof:

wherein A, B, R₁, R₃, R₄, n, and v are as previously defined.

In another embodiment, the invention provides a compound represented byone of Formulas (IVa) (IVc) or a pharmaceutically acceptable salt, esteror prodrug thereof:

wherein A, B, R₁, R₃, and R₄ are as previously defined.

In another embodiment, the invention provides a compound represented byone of Formulas (Va)˜(Vc) or a pharmaceutically acceptable salt, esteror prodrug thereof:

wherein A, B, R₃, and R₄ are as previously defined.

In certain embodiments of the compounds of Formulas (I), (IIa)-(IIc),(IIIa)-(IIIc), (IVa)-(IVc) and (Va)-(Vc), A is derived from one of thefollowing by removal of two hydrogen atoms:

wherein each of the above is optionally substituted when possible.Preferably, when present, the substituents are independently selectedfrom, but not limited to, —CN, —F, —Cl, —CH₃, —CF₃, —OCH₃, and —OCF₃.

In certain embodiments of the compounds of Formulas (I), (IIa)-(IIc),(IIIa)-(IIIc), (IVa)-(IVc) and (Va)-(Vc), A is selected from thefollowing:

wherein each of the above is optionally substituted. Preferably, whenpresent, the substituents are independently selected from, but notlimited to, —CN, —F, —Cl, —CH₃, —CF₃, —OCH₃, and —OCF₃.

In another embodiment, the invention provides a compound represented byone of Formulas (VIa)˜(VIc) or a pharmaceutically acceptable salt, esteror prodrug thereof:

wherein B, R₁, R₃, and R₄ are as previously defined.

In another embodiment, the invention provides a compound represented byone of Formulas (VIIa)˜(VIIc) or a pharmaceutically acceptable salt,ester or prodrug thereof:

wherein B, R₃, and R₄ are as previously defined.

In certain embodiments of the compounds of Formulas (I), (IIa)-(IIc),(IIIa)-(IIIc), (IVa)-(IVc), (Va)-(Vc), (VIa)-(VIc) and (VIIa)-(VIIc), Bis derived from one of the following by removal of a ring hydrogen atom:

wherein each of the above is optionally substituted when possible.Preferably, when present, the substituents are independently selectedfrom, but not limited to, —CN, —F, —Cl, —CH₃, —CF₃, —OCH₃, and —OCF₃.

It will be appreciated that the description of the present inventionherein should be construed in congruity with the laws and principles ofchemical bonding. In some instances it may be necessary to remove ahydrogen atom in order to accommodate a substituent at any givenlocation.

It is intended that the definition of any substituent or variable (e.g.,R₁, R₂, etc.) at a particular location in a molecule be independent ofits definitions elsewhere in that molecule.

It will be yet appreciated that the compounds of the present inventionmay contain one or more asymmetric carbon atoms and may exist inracemic, diastereoisomeric, and optically active forms. It will still beappreciated that certain compounds of the present invention may exist indifferent tautomeric forms. All tautomers are contemplated to be withinthe scope of the present invention.

In certain embodiments, the present invention provides a method for theprevention or treatment of RSV activities and for treating RSV infectionin a subject in need thereof. The method comprises administering to thesubject a therapeutically effective amount of a compound of Formula (I).

The present invention also provides the use of a compound of Formula (I)for the preparation of a medicament for the prevention or treatment ofRSV.

Thus, in one embodiment, a compound of Formula (I), or pharmaceuticallyacceptable salt thereof, is combined with a steroid anti-inflammatorycompound, for example budesonide or fluticasone. In a preferredembodiment, the steroid is administered in low doses to minimizeimmuno-suppressant effects. In another embodiment a compound of Formula(I), or a pharmaceutically acceptable salt thereof, is combined with anon-steroid anti-inflammatory compound, for example leukotrieneantagonists such as Singulair (Merck) or Accolate (Astra Zeneca),phosphodiesterase 4 inhibitors such as roflumilast (Altana), TNF alphainhibitors such as Enbrel (Amgen), Remicade (Centocor), Humira (Abbott)or CDP870 (Celltech) or NSAIDS. In a further embodiment, a compound ofFormula (I) is combined with interleukin 8 or interleukin 9 inhibitors.The present invention thus also relates to a product containing acompound of Formula (I), or a pharmaceutically acceptable salt thereof,and an anti-inflammatory compound for simultaneous, separate orsequential use in the treatment of RSV.

The present invention also relates to a combination of a compound ofFormula (I), or a pharmaceutically acceptable salt thereof, with ananti-influenza compound and the use of such a combination in thetreatment of concomitant RSVand influenza infections. The presentinvention thus also relates to a product containing a compound ofFormula (I), or a pharmaceutically acceptable salt thereof, and ananti-influenza compound for simultaneous, separate or sequential use inthe treatment of concomitant RSV and influenza infections. The compoundsof the invention may be administered in a variety of dosage forms. Thus,they can be administered orally, for example as tablets, troches,lozenges, aqueous or oily suspensions, dispersible powders or granules.The compounds of the invention may also be administered parenterally,whether subcutaneously, intravenously, intramuscularly, intrasternally,transdermally or by infusion techniques. The compounds may also beadministered as suppositories.

In an embodiment, the compounds of the invention are administered byintranasal or intrabronchial administration. The present invention alsoprovides an inhaler or nebuliser containing a medicament which comprises(a) a benzodiazepine derivative of the Formula (I), as defined above, ora pharmaceutically acceptable salt thereof, and (b) a pharmaceuticallyacceptable carrier or diluent.

The present invention also provides a pharmaceutical compositioncontaining such a benzodiazepine derivative, or a pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier ordiluent.

The compounds of the invention are typically formulated foradministration with a pharmaceutically acceptable carrier or diluent.For example, solid oral forms may contain, together with the activecompound, diluents, e.g. lactose, dextrose, saccharose, cellulose, cornstarch or potato starch; lubricants, e.g. silica, talc, stearic acid,magnesium or calcium stearate, and/or polyethylene glycols; bindingagents; e.g. starches, arabic gums, gelatin, methylcellulose,carboxymethylcellulose or polyvinyl pyrrolidone; disaggregating agents,e.g. starch, alginic acid, alginates or sodium starch glycolate;effervescing mixtures; dyestuffs; sweeteners; wetting agents, such aslecithin, polysorbates, laurylsulphates; and, in general, non toxic andpharmacologically inactive substances used in pharmaceuticalformulations. Such pharmaceutical preparations may be manufactured inknown manner, for example, by means of mixing, granulating, tableting,sugar coating, or film coating processes.

Liquid dispersions for oral administration may be syrups, emulsions andsuspensions. The syrups may contain as carriers, for example, saccharoseor saccharose with glycerine and/or mannitol and/or sorbitol.

Suspensions and emulsions may contain as carrier, for example a naturalgum, agar, sodium alginate, pectin, methylcellulose,carboxymethylcellulose, or polyvinyl alcohol. The suspension orsolutions for intramuscular injections may contain, together with theactive compound, a pharmaceutically acceptable carrier, e.g., sterilewater, olive oil, ethyl oleate, glycols, e.g. propylene glycol, and ifdesired, a suitable amount of lidocaine hydrochloride.

Solutions for injection or infusion may contain as carrier, for example,sterile water or preferably they may be in the form of sterile, aqueous,isotonic saline solutions.

The present invention also relates to the novel compounds, as definedabove; or a pharmaceutically acceptable salt thereof, for use in amethod of treating the human or animal body. The present invention alsorelates to a pharmaceutical composition comprising a novel compound asdefined above and a pharmaceutically acceptable diluant or carrier.Preferably, the pharmaceutical composition comprises a pharmaceuticallyacceptable salt of a novel compound as defined above. A pharmaceuticallyacceptable salt is as defined above. The novel compounds of theinvention are typically administered in the manner defined above and thecompounds are typically formulated for administration in the mannerdefined above.

Preferably, the pharmaceutical compositions comprise optically activeisomers of the novel compounds of the invention. Thus, for example,preferred novel compounds of the invention containing only one chiralcentre include an R enantiomer in substantially pure form, an Senantiomer in substantially pure form and enantiomeric mixtures whichcontain an excess of the R enantiomer or an excess of the S enantiomer.It is particularly preferred that pharmaceutical contains a compound ofthe invention which is a substantially pure optical isomer. For theavoidance of doubt, the novel compounds of the invention can, ifdesired, be used in the form of solvates.

Yet a further aspect of the present invention is a process of making anyof the compounds delineated herein employing any of the synthetic meansdelineated herein.

Definitions

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

The term “aryl,” as used herein, refers to a mono-, bi-, or polycycliccarbocyclic ring system comprising at least one aromatic ring,including, but not limited to, phenyl, naphthyl, tetrahydronaphthyl,indanyl, and indenyl. A polycyclic aryl is a polycyclic ring system thatcomprises at least one aromatic ring. Polycyclic aryls can comprisefused rings, covalently attached rings or a combination thereof.

The term “heteroaryl,” as used herein, refers to a mono-, bi-, orpolycyclic aromatic radical having one or more ring atom selected fromS, O and N; and the remaining ring atoms are carbon, wherein any N or Scontained within the ring may be optionally oxidized. Heteroarylincludes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl,pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl,thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl,isoquinolinyl, benzimidazolyl, benzoxazolyl, quinoxalinyl. A polycyclicheteroaryl can comprise fused rings, covalently attached rings or acombination thereof.

In accordance with the invention, aromatic groups can be substituted orunsubstituted.

The term “bicyclic aryl” or “bicyclic heteroaryl” refers to a ringsystem consisting of two rings wherein at least one ring is aromatic;and the two rings can be fused or covalently attached.

The term “alkyl” as used herein, refers to saturated, straight- orbranched-chain hydrocarbon radicals. “C₁-C₃ alkyl,” “C₁-C₆ alkyl,”“C₁-C₁₀ alkyl” C₂-C₄ alkyl,” or “C₃-C₆ alkyl,” refer to alkyl groupscontaining from one to three, one to six, one to ten carbon atoms, 2 to4 and 3 to 6 carbon atoms respectively. Examples of C₁-C₅ alkyl radicalsinclude, but are not limited to, methyl, ethyl, propyl, isopropyl,n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl and octyl radicals.

The term “alkenyl” as used herein, refers to straight- or branched-chainhydrocarbon radicals having at least one carbon-carbon double bond bythe removal of a single hydrogen atom. “C₂-C₁₀ alkenyl,” “C₂-C₈alkenyl,” “C₂-C₄ alkenyl,” or “C₃-C₆ alkenyl,” refer to alkenyl groupscontaining from two to ten, two to eight, two to four or three to sixcarbon atoms respectively. Alkenyl groups include, but are not limitedto, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl,heptenyl, octenyl, and the like.

The term “alkynyl” as used herein, refers to straight- or branched-chainhydrocarbon radicals having at least one carbon-carbon triple bond bythe removal of a single hydrogen atom. “C₂-C₁₀ alkynyl,” “C₂-C₈alkynyl,” “C₂-C₄ alkynyl,” or “C₃-C₆ alkynyl,” refer to alkynyl groupscontaining from two to ten, two to eight, two to four or three to sixcarbon atoms respectively. Representative alkynyl groups include, butare not limited to, for example, ethynyl, 1-propynyl, 1-butynyl,heptynyl, octynyl, and the like.

The term “cycloalkyl”, as used herein, refers to a monocyclic orpolycyclic saturated carbocyclic ring or a bi- or tri-cyclic groupfused, bridged or spiro system, and the carbon atoms may be optionallyoxo-substituted or optionally substituted with exocyclic olefinic,iminic or oximic double bond. Preferred cycloalkyl groups include C₃-C₁₂cycloalkyl, C₃-C₆ cycloalkyl, C₃-C₈ cycloalkyl and C₄-C₇ cycloalkyl.Examples of C₃-C₁₂ cycloalkyl include, but not limited to, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl,4-methylene-cyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl,spiro[2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, spiro[4.4]nonanyl, andthe like.

The term “cycloalkenyl”, as used herein, refers to monocyclic orpolycyclic carbocyclic ring or a bi- or tri-cyclic group fused, bridgedor spiro system having at least one carbon-carbon double bond and thecarbon atoms may be optionally oxo-substituted or optionally substitutedwith exocyclic olefinic, iminic or oximic double bond. Preferredcycloalkenyl groups include C₃-C₁₂ cycloalkenyl, C₃-C₈ cycloalkenyl orC₅-C₇ cycloalkenyl groups. Examples of C₃-C₁₂ cycloalkenyl include, butnot limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, cyclooctenyl, bicyclo[2.2.1]hept-2-enyl,bicyclo[3.1.0]hex-2-enyl, spiro[2.5]oct-4-enyl, spiro[4.4]non-1-enyl,bicyclo[4.2.1]non-3-en-9-yl, and the like.

As used herein, the term “arylalkyl” means a functional group wherein analkylene chain is attached to an aryl group, e.g., —CH₂CH₂-phenyl. Theterm “substituted arylalkyl” means an arylalkyl functional group inwhich the aryl group is substituted. Similarly, the term“heteroarylalkyl” means a functional group wherein an alkylene chain isattached to a heteroaryl group. The term “substituted heteroarylalkyl”means a heteroarylalkyl functional group in which the heteroaryl groupis substituted.

As used herein, the term “alkoxy” employed alone or in combination withother terms means, unless otherwise stated, an alkyl group having thedesignated number of carbon atoms connected to the rest of the moleculevia an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy,2-propoxy (isopropoxy) and the higher homologs and isomers. Preferredalkoxy are (C₁-C₃) alkoxy.

It is understood that any alkyl, alkenyl, alkynyl, cycloalkyl,heterocyclic and cycloalkenyl moiety described herein can also be analiphatic group or an alicyclic group.

An “aliphatic” group is a non-aromatic moiety comprised of anycombination of carbon atoms, hydrogen atoms, halogen atoms, oxygen,nitrogen or other atoms, and optionally contains one or more units ofunsaturation, e.g., double and/or triple bonds. Examples of aliphaticgroups are functional groups, such as alkyl, alkenyl, alkynyl, O, OH,NH, NH₂, C(O), S(O)₂, C(O)O, C(O)NH, OC(O)O, OC(O)NH, OC(O)NH₂, S(O)₂NH,S(O)₂NH₂, NHC(O)NH₂, NHC(O)C(O)NH, NHS(O)₂NH, NHS(O)₂NH₂, C(O)NHS(O)₂,C(O)NHS(O)₂NH or C(O)NHS(O)₂NH₂, and the like, groups comprising one ormore functional groups, non-aromatic hydrocarbons (optionallysubstituted), and groups wherein one or more carbons of a non-aromatichydrocarbon (optionally substituted) is replaced by a functional group.Carbon atoms of an aliphatic group can be optionally oxo-substituted. Analiphatic group may be straight chained, branched, cyclic, or acombination thereof and preferably contains between about 1 and about 24carbon atoms, more typically between about 1 and about 12 carbon atoms.In addition to aliphatic hydrocarbon groups, as used herein, aliphaticgroups expressly include, for example, alkoxyalkyls, polyalkoxyalkyls,such as polyalkylene glycols, polyamines, and polyimines, for example.Aliphatic groups may be optionally substituted.

The terms “heterocyclic” or “heterocycloalkyl” can be usedinterchangeably and referred to a non-aromatic ring or a bi- ortri-cyclic group fused, bridged or spiro system, where (i) each ringsystem contains at least one heteroatom independently selected fromoxygen, sulfur and nitrogen, (ii) each ring system can be saturated orunsaturated (iii) the nitrogen and sulfur heteroatoms may optionally beoxidized, (iv) the nitrogen heteroatom may optionally be quaternized,(v) any of the above rings may be fused to an aromatic ring, and (vi)the remaining ring atoms are carbon atoms which may be optionallyoxo-substituted or optionally substituted with exocyclic olefinic,iminic or oximic double bond. Representative heterocycloalkyl groupsinclude, but are not limited to, 1,3-dioxolane, pyrrolidinyl,pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl,piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl,isothiazolidinyl, quinoxalinyl, pyridazinonyl,2-azabicyclo[2.2.1]-heptyl, 8-azabicyclo[3.2.1]octyl,5-azaspiro[2.5]octyl, 1-oxa-7-azaspiro[4.4]nonanyl, 7-oxooxepan-4-yl,and tetrahydrofuryl. Such heterocyclic groups may be furthersubstituted. Heteroaryl or heterocyclic groups can be C-attached orN-attached (where possible).

It is understood that any alkyl, alkenyl, alkynyl, alicyclic,cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclic, aliphaticmoiety or the like, described herein can also be a divalent ormultivalent group when used as a linkage to connect two or more groupsor substituents, which can be at the same or different atom(s). One ofskill in the art can readily determine the valence of any such groupfrom the context in which it occurs.

The term “substituted” refers to substitution by independent replacementof one, two, or three or more of the hydrogen atoms with substituentsincluding, but not limited to, —F, —Cl, —Br, —I, —OH, C₁-C₁₂-alkyl;C₂-C₁₂-alkenyl, C₂-C₁₂-alkynyl, —C₃-C₂-cycloalkyl, protected hydroxy,—NO₂, —N₃, —CN, —NH₂, protected amino, oxo, thioxo, —NH—C₁-C₁₂-alkyl,—NH—C₂-C₈-alkenyl, —NH—C₂-C₈-alkynyl, —NH—C₃-C₂-cycloalkyl, —NH-aryl,—NH-heteroaryl, —NH-heterocycloalkyl, -dialkylamino, -diarylamino,-diheteroarylamino, —O—C₁-C₁₂-alkyl, —O—C₂-C₈-alkenyl, —O—C₂-C₈-alkynyl,—O—C₃-C₁₂-cycloalkyl, —O-aryl, —O-heteroaryl, —O-heterocycloalkyl,—C(O)—C₁-C₁₂-alkyl, —C(O)—C₂-C₈-alkenyl, —C(O)—C₂-C₈-alkynyl,—C(O)—C₃-C₁₂-cycloalkyl, —C(O)-aryl, —C(O)— heteroaryl,—C(O)-heterocycloalkyl, —CONH₂, —CONH—C₁-C₁₂-alkyl, —CONH—C₂-C₈-alkenyl,—CONH—C₂-C₈-alkynyl, —CONH—C₃-C₁₂-cycloalkyl, —CONH-aryl,—CONH-heteroaryl, —CONH— heterocycloalkyl, —OCO₂—C₁-C₁₂-alkyl,—OCO₂—C₂-C₈-alkenyl, —OCO₂—C₂-C₈-alkynyl, —OCO₂—C₃-C₁₂-cycloalkyl,—OCO₂-aryl, —OCO₂-heteroaryl, —OCO₂-heterocycloalkyl, —CO₂—C₁-C₁₂ alkyl,—CO₂—C₂-C₈ alkenyl, —CO₂—C₂-C₈ alkynyl, CO₂—C₃-C₁₂-cycloalkyl, —CO₂—aryl, CO₂-heteroaryl, CO₂-heterocyloalkyl, —OCONH₂, —OCONH—C₁-C₁₂-alkyl,—OCONH—C₂-C₈-alkenyl, —OCONH—C₂-C₈-alkynyl, —OCONH—C₃-C₁₂-cycloalkyl,—OCONH-aryl, —OCONH-heteroaryl, —OCONH-heterocyclo-alkyl, —NHC(O)H,—NHC(O)—C₁-C₁₂-alkyl, —NHC(O)—C₂-C₈-alkenyl, —NHC(O)—C₂-C₈-alkynyl,—NHC(O)—C₃-C₂-cycloalkyl, —NHC(O)-aryl, —NHC(O)-heteroaryl,—NHC(O)-heterocyclo-alkyl, —NHCO₂—C₁-C₁₂-alkyl, —NHCO₂—C₂-C₈-alkenyl,—NHCO₂— C₂-C₈-alkynyl, —NHCO₂—C₃-C₁₂-cycloalkyl, —NHCO₂-aryl,—NHCO₂-heteroaryl, —NHCO₂— heterocycloalkyl, —NHC(O)NH₂,—NHC(O)NH—C₁-C₁₂-alkyl, —NHC(O)NH—C₂-C₈-alkenyl,—NHC(O)NH—C₂-C₈-alkynyl, —NHC(O)NH—C₃-C₁₂-cycloalkyl, —NHC(O)NH-aryl,—NHC(O)NH-heteroaryl, —NHC(O)NH-heterocycloalkyl, NHC(S)NH₂,—NHC(S)NH—C₁-C₁₂-alkyl, —NHC(S)NH—C₂-C₈-alkenyl,—NHC(S)NH—C₂-C₈-alkynyl, —NHC(S)NH—C₃-C₁₂-cycloalkyl, —NHC(S)NH-aryl,—NHC(S)NH-heteroaryl, —NHC(S)NH-heterocycloalkyl, —NHC(NH)NH₂,—NHC(NH)NH—C₁-C₁₂-alkyl, —NHC(NH)NH—C₂-C₈-alkenyl,—NHC(NH)NH—C₂-C₈-alkynyl, —NHC(NH)NH—C₃-C₁₂-cycloalkyl, —NHC(NH)NH-aryl,—NHC(NH)NH-heteroaryl, —NHC(NH)NH-heterocycloalkyl,—NHC(NH)—C₁-C₁₂-alkyl, —NHC(NH)—C₂-C₈-alkenyl, —NHC(NH)—C₂-C₈-alkynyl,—NHC(NH)—C₃-C₁₂-cycloalkyl, —NHC(NH)-aryl, —NHC(NH)-heteroaryl,—NHC(NH)-heterocycloalkyl, —C(NH)NH—C₁-C₁₂-alkyl,—C(NH)NH—C₂-C₈-alkenyl, —C(NH)NH—C₂-C₈-alkynyl,—C(NH)NH—C₃-C₁₂-cycloalkyl, —C(NH)NH-aryl, —C(NH)NH-heteroaryl,—C(NH)NH-heterocycloalkyl, —S(O)—C₁-C₁₂-alkyl, —S(O)—C₂-C₈-alkenyl,—S(O)—C₂-C₈-alkynyl, —S(O)—C₃-C₁₂-cycloalkyl, —S(O)-aryl,—S(O)-heteroaryl, —S(O)-heterocycloalkyl, —SO₂NH₂, —SO₂NH—C₁-C₁₂-alkyl,—SO₂NH—C₂-C₈-alkenyl, —SO₂NH—C₂-C₈-alkynyl, —SO₂NH—C₃-C₁₂-cycloalkyl,—SO₂NH-aryl, —SO₂NH-heteroaryl, —SO₂NH-heterocycloalkyl,—NHSO₂—C₁-C₁₂-alkyl, —NHSO₂—C₂-C₈-alkenyl, —NHSO₂—C₂-C₈-alkynyl,—NHSO₂—C₃-C₁₂-cycloalkyl, —NHSO₂-aryl, —NHSO₂-heteroaryl, —NHSO₂—heterocycloalkyl, —CH₂NH₂, —CH₂SO₂CH₃, -aryl, -arylalkyl, -heteroaryl,-heteroarylalkyl, -heterocycloalkyl, —C₃-C₁₂-cycloalkyl,polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, —SH,—S—C₁-C₁₂-alkyl, —S—C₂-C₈-alkenyl, —S—C₂-C₈-alkynyl,—S—C₃-C₁₂-cycloalkyl, —S-aryl, —S-heteroaryl, —S-heterocycloalkyl, ormethylthio-methyl. In certain embodiments, the substituents areindependently selected from halo, preferably Cl and F; C₁-C₄-alkyl,preferably methyl and ethyl; halo-C₁-C₄-alkyl, such as fluoromethyl,difluoromethyl, and trifluoromethyl; C₂-C₄-alkenyl; halo-C₂-C₄-alkenyl;C₃-C₆-cycloalkyl, such as cyclopropyl; C₁-C₄-alkoxy, such as methoxy andethoxy; halo-C₁-C₄-alkoxy, such as fluoromethoxy, difluoromethoxy, andtrifluoromethoxy, —CN; —OH; NH₂; C₁-C₄-alkylamino; di(C₁-C₄-alkyl)amino;and NO₂. It is understood that the aryls, heteroaryls, alkyls, and thelike can be further substituted. In some cases, each substituent in asubstituted moiety is additionally optionally substituted when possiblewith one or more groups, each group being independently selected fromC₁-C₄-alkyl; —CF₃, —OCH₃, —OCF₃, —F, —Cl, —Br, —I, —OH, —NO₂, —CN, and—NH₂. Preferably, a substituted alkyl group, such as a substitutedmethyl group, is substituted with one or more halogen atoms, morepreferably one or more fluorine or chlorine atoms.

In certain embodiments, a substituted alkyl, alkenyl or alkoxy group issubstituted with one or more halogen atoms, preferably fluorine atoms.Such substituted alkyl groups include fluoromethyl, difluoromethyl andtrifluoromethyl. Such substituted alkoxy groups include fluoromethoxy,difluoromethoxy and trifluoromethoxy.

The term “halo” or halogen” alone or as part of another substituent, asused herein, refers to a fluorine, chlorine, bromine, or iodine atom.

The term “optionally substituted”, as used herein, means that thereferenced group may be substituted or unsubstituted. In one embodiment,the referenced group is optionally substituted with zero substituents,i.e., the referenced group is unsubstituted. In another embodiment, thereferenced group is optionally substituted with one or more additionalgroup(s) individually and independently selected from groups describedherein.

The term “hydrogen” includes hydrogen and deuterium. In addition, therecitation of an atom includes other isotopes of that atom so long asthe resulting compound is pharmaceutically acceptable.

In certain embodiments, the compounds of each formula herein are definedto include isotopically labelled compounds. An “isotopically labelledcompound” is a compound in which at least one atomic position isenriched in a specific isotope of the designated element to a levelwhich is significantly greater than the natural abundance of thatisotope. For example, one or more hydrogen atom positions in a compoundcan be enriched with deuterium to a level which is significantly greaterthan the natural abundance of deuterium, for example, enrichment to alevel of at least 1%, preferably at least 20% or at least 50%. Such adeuterated compound may, for example, be metabolized more slowly thanits non-deuterated analog, and therefore exhibit a longer half-life whenadministered to a subject. Such compounds can synthesize using methodsknown in the art, for example by employing deuterated startingmaterials. Unless stated to the contrary, isotopically labelledcompounds are pharmaceutically acceptable.

The term “hydroxy activating group,” as used herein, refers to a labilechemical moiety which is known in the art to activate a hydroxyl groupso that it will depart during synthetic procedures such as in asubstitution or an elimination reaction. Examples of hydroxyl activatinggroup include, but are not limited to, mesylate, tosylate, triflate,p-nitrobenzoate, phosphonate and the like.

The term “activated hydroxyl,” as used herein, refers to a hydroxy groupactivated with a hydroxyl activating group, as defined above, includingmesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups, forexample.

The term “hydroxy protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect a hydroxyl groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the hydroxy protecting group as described hereinmay be selectively removed. Hydroxy protecting groups as known in theart are described generally in T. H. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons,New York (1999). Examples of hydroxyl protecting groups includebenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxy-carbonyl,isopropoxycarbonyl, diphenylmethoxycarbonyl,2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl,chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl,methyl, t-butyl, 2,2,2-trichloroethyl, 2-trimethylsilyl ethyl, allyl,benzyl, triphenyl-methyl (trityl), methoxymethyl, methylthiomethyl,benzyloxymethyl, 2-(trimethylsilyl)-ethoxymethyl, methanesulfonyl,trimethylsilyl, triisopropylsilyl, and the like.

The term “protected hydroxy,” as used herein, refers to a hydroxy groupprotected with a hydroxy protecting group, as defined above, includingbenzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl groups,for example.

The term “hydroxy prodrug group,” as used herein, refers to a promoietygroup which is known in the art to change the physicochemical, and hencethe biological properties of a parent drug in a transient manner bycovering or masking the hydroxy group. After said syntheticprocedure(s), the hydroxy prodrug group as described herein must becapable of reverting back to hydroxy group in vivo. Hydroxy prodruggroups as known in the art are described generally in Kenneth B. Sloan,Prodrugs, Topical and Ocular Drug Delivery, (Drugs and thePharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York(1992) and in “Prodrugs of Alcohols and Phenols” by S. S. Dhareshwar andV. J. Stella, in Prodrugs Challenges and Rewards Part-2, (Biotechnology:Pharmaceutical Aspects), edited by V. J. Stella, et al, Springer andAAPSPress, 2007, pp 31-99.

The term “amino protecting group,” as used herein, refers to a labilechemical moiety which is known in the art to protect an amino groupagainst undesired reactions during synthetic procedures. After saidsynthetic procedure(s) the amino protecting group as described hereinmay be selectively removed. Amino protecting groups as known in the artare described generally in T. H. Greene and P. G. M. Wuts, ProtectiveGroups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York(1999). Examples of amino protecting groups include, but are not limitedto, methoxycarbonyl, t-butoxycarbonyl, 9-fluorenyl-methoxycarbonyl,benzyloxycarbonyl, and the like.

The term “protected amino,” as used herein, refers to an amino groupprotected with an amino protecting group as defined above.

The term “leaving group” means a functional group or atom which can bedisplaced by another functional group or atom in a substitutionreaction, such as a nucleophilic substitution reaction. By way ofexample, representative leaving groups include chloro, bromo and iodogroups; sulfonic ester groups, such as mesylate, tosylate, brosylate,nosylate and the like; and acyloxy groups, such as acetoxy,trifluoroacetoxy and the like.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such compounds are well known to those skilledin the art, and it will be obvious to those skilled in the art thatindividual solvents or mixtures thereof may be preferred for specificcompounds and reaction conditions, depending upon such factors as thesolubility of reagents, reactivity of reagents and preferred temperatureranges, for example. Further discussions of aprotic solvents may befound in organic chemistry textbooks or in specialized monographs, forexample: Organic Solvents Physical Properties and Methods ofPurification, 4th ed., edited by John A. Riddick et al., Vol. II, in theTechniques of Chemistry Series, John Wiley & Sons, N Y, 1986.

The term “protic solvent,” as used herein, refers to a solvent thattends to provide protons, such as an alcohol, for example, methanol,ethanol, propanol, isopropanol, butanol, t-butanol, and the like. Suchsolvents are well known to those skilled in the art, and it will beobvious to those skilled in the art that individual solvents or mixturesthereof may be preferred for specific compounds and reaction conditions,depending upon such factors as the solubility of reagents, reactivity ofreagents and preferred temperature ranges, for example. Furtherdiscussions of protogenic solvents may be found in organic chemistrytextbooks or in specialized monographs, for example: Organic SolventsPhysical Properties and Methods of Purification, 4th ed., edited by JohnA. Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, N Y, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable,” as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the Formula herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, 2^(nd) Ed. Wiley-VCH (1999); T. W. Greene and P. G. M.Wuts, Protective Groups in Organic Synthesis, 3rd Ed., John Wiley andSons (1999); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The term “subject,” as used herein, refers to an animal. Preferably, theanimal is a mammal. More preferably, the mammal is a human. A subjectalso refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, fish, birds and the like.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and may include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds, otherunsaturation, or other centers of geometric asymmetry, and unlessspecified otherwise, it is intended that the compounds include both Eand Z geometric isomers or cis- and trans-isomers. Likewise, alltautomeric forms are also intended to be included. Tautomers may be incyclic or acyclic. The configuration of any carbon-carbon double bondappearing herein is selected for convenience only and is not intended todesignate a particular configuration unless the text so states; thus acarbon-carbon double bond or carbon-heteroatom double bond depictedarbitrarily herein as trans may be cis, trans, or a mixture of the twoin any proportion.

Certain compounds of the present invention may also exist in differentstable conformational forms which may be separable. Torsional asymmetrydue to restricted rotation about an asymmetric single bond, for examplebecause of steric hindrance or ring strain, may permit separation ofdifferent conformers. The present invention includes each conformationalisomer of these compounds and mixtures thereof.

As used herein, the term “pharmaceutically acceptable salt,” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid. Examples of pharmaceutically acceptable saltsinclude, but are not limited to, nontoxic acid addition salts are saltsof an amino group formed with inorganic acids such as hydrochloric acid,hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid orwith organic acids such as acetic acid, maleic acid, tartaric acid,citric acid, succinic acid or malonic acid or by using other methodsused in the art such as ion exchange. Other pharmaceutically acceptablesalts include, but are not limited to, adipate, alginate, ascorbate,aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate,camphorate, camphorsulfonate, citrate, cyclopentane-propionate,digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate,glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate,hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate,lactate, laurate, lauryl sulfate, malate, maleate, malonate,methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate,oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate,phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate,tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts,and the like. Representative alkali or alkaline earth metal saltsinclude sodium, lithium, potassium, calcium, magnesium, and the like.Further pharmaceutically acceptable salts include, when appropriate,nontoxic ammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

Pharmaceutically acceptable salts can also be prepared by deprotonationof the parent compound with a suitable base, thereby forming the anionicconjugate base of the parent compound. In such salts the counter ion isa cation. Suitable cations include ammonium and metal cations, such asalkali metal cations, including Li⁺, Na⁺, K⁺ and Cs⁺, and alkaline earthmetal cations, such as Mg²⁺ and Ca²⁺.

As used herein, the term “pharmaceutically acceptable ester” refers toesters which hydrolyze in vivo and include those that break down readilyin the human body to leave the parent compound or a salt thereof.Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Examples of particular esters include, but are not limited to, esters ofC₁-C₆-alkanoic acids, such as acetate, propionate, butyrate and pivalateesters.

In certain embodiments, the invention provides pharmaceuticallyacceptable prodrugs of the compounds disclosed herein. The term“pharmaceutically acceptable prodrugs” as used herein refers to thoseprodrugs of the compounds formed by the process of the present inventionwhich are, within the scope of sound medical judgment, suitable for usein contact with the tissues of humans and lower animals with unduetoxicity, irritation, allergic response, and the like, commensurate witha reasonable benefit/risk ratio, and effective for their intended use,as well as the zwitterionic forms, where possible, of the compounds ofthe present invention. “Prodrug”, as used herein means a compound, whichis convertible in vivo by metabolic means (e.g. by hydrolysis) to affordany compound delineated by the formulae of the instant invention.Various forms of prodrugs are known in the art, for example, asdiscussed in Bundgaard, (ed.), Design of Prodrugs, Elsevier (1985);Widder, et al. (ed.), Methods in Enzymology, Vol. 4, Academic Press(1985); Krogsgaard-Larsen, et al., (ed.). “Design and Application ofProdrugs, Textbook of Drug Design and Development, Chapter 5, 113-191(1991); Bundgaard, et al., Journal of Drug Deliver Reviews,8:1-38(1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq.(1988); Higuchi and Stella (eds.) Prodrugs as Novel Drug DeliverySystems, American Chemical Society (1975); and Bernard Testa & JoachimMayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry,Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

Additional types of prodrugs are also encompassed. For instance, freecarboxyl groups can be derivatized as amides or alkyl esters. Freehydroxy groups may be derivatized using groups including but not limitedto hemisuccinates, ethyl succinate, phosphate esters,dimethylaminoacetates, and phosphoryloxymethyloxycarbonyls, as outlinedin Advanced Drug Delivery Reviews, 1996, 19, 115. Carbamate prodrugs ofhydroxy and amino groups are also included, as are carbonate prodrugs,sulfonate esters and sulfate esters of hydroxy groups. Derivatization ofhydroxy groups as (acyloxy)methyl and (acyloxy)ethyl ethers wherein theacyl group may be an alkyl ester, optionally substituted with groupsincluding but not limited to ether, amine and carboxylic acidfunctionalities, or where the acyl group is an amino acid ester asdescribed above, are also encompassed. Prodrugs of this type aredescribed in J. Med. Chem. 1996, 39, 10. Free amines can also bederivatized as amides, sulfonamides or phosphonamides. All of theseprodrug moieties may incorporate groups including but not limited toether, amine and carboxylic acid functionalities. In certainembodiments, a compound of the invention can incorporate two or moregroups that are metabolically removed in vivo to yield the active parentcompound. For example, a compound of Formula I wherein R₁ is an aminoacid residue can also be esterified, for example at a hydroxyl group ofthe sugar residue, to form a compound with two groups that can beremoved in vivo to yield the active compound.

The term “treating”, as used herein, means relieving, lessening,reducing, eliminating, modulating, or ameliorating, i.e. causingregression of the disease state or condition. Treating can also includeinhibiting, i.e. arresting the development, of a existing disease stateor condition, and relieving or ameliorating, i.e. causing regression ofan existing disease state or condition, for example when the diseasestate or condition may already be present.

The term “preventing”, as used herein means, to completely or almostcompletely stop a disease state or condition, from occurring in apatient or subject, especially when the patient or subject ispredisposed to such or at risk of contracting a disease state orcondition.

Additionally, the compounds of the present invention, for example, thesalts of the compounds, can exist in either hydrated or unhydrated (theanhydrous) form or as solvates with other solvent molecules. Nonlimitingexamples of hydrates include monohydrates, dihydrates, etc. Nonlimitingexamples of solvates include ethanol solvates, acetone solvates, etc.“Solvates” means solvent addition forms that contain eitherstoichiometric or non stoichiometric amounts of solvent. Some compoundshave a tendency to trap a fixed molar ratio of solvent molecules in thecrystalline solid state, thus forming a solvate. If the solvent is waterthe solvate formed is a hydrate, when the solvent is alcohol, thesolvate formed is an alcoholate. Hydrates are formed by the combinationof one or more molecules of water with one of the substances in whichthe water retains its molecular state as H₂O, such combination beingable to form one or more hydrate.

As used herein, the term “analog” refers to a chemical compound that isstructurally similar to another but differs slightly in composition (asin the replacement of one atom by an atom of a different element or inthe presence of a particular functional group, or the replacement of onefunctional group by another functional group). Thus, an analog is acompound that is similar to or comparable in function and appearance tothe reference compound.

The term “aprotic solvent,” as used herein, refers to a solvent that isrelatively inert to proton activity, i.e., not acting as a proton-donor.Examples include, but are not limited to, hydrocarbons, such as hexaneand toluene, for example, halogenated hydrocarbons, such as, forexample, methylene chloride, ethylene chloride, chloroform, and thelike, heterocyclic compounds, such as, for example, tetrahydrofuran andN-methylpyrrolidinone, and ethers such as diethyl ether,bis-methoxymethyl ether. Such solvents are well known to those skilledin the art, and individual solvents or mixtures thereof may be preferredfor specific compounds and reaction conditions, depending upon suchfactors as the solubility of reagents, reactivity of reagents andpreferred temperature ranges, for example. Further discussions ofaprotic solvents may be found in organic chemistry textbooks or inspecialized monographs, for example: Organic Solvents PhysicalProperties and Methods of Purification, 4th ed., edited by John A.Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, N Y, 1986.

The terms “protogenic organic solvent” or “protic solvent” as usedherein, refer to a solvent that tends to provide protons, such as analcohol, for example, methanol, ethanol, propanol, isopropanol, butanol,t-butanol, and the like. Such solvents are well known to those skilledin the art, and individual solvents or mixtures thereof may be preferredfor specific compounds and reaction conditions, depending upon suchfactors as the solubility of reagents, reactivity of reagents andpreferred temperature ranges, for example. Further discussions ofprotogenic solvents may be found in organic chemistry textbooks or inspecialized monographs, for example: Organic Solvents PhysicalProperties and Methods of Purification, 4th ed., edited by John A.Riddick et al., Vol. II, in the Techniques of Chemistry Series, JohnWiley & Sons, N Y, 1986.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., therapeutic or prophylacticadministration to a subject).

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. Additionally, thevarious synthetic steps may be performed in an alternate sequence ororder to give the desired compounds. In addition, the solvents,temperatures, reaction durations, etc. delineated herein are forpurposes of illustration only and variation of the reaction conditionscan produce the desired bridged macrocyclic products of the presentinvention. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein include, for example, those described in R.Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 2d.Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser andFieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); andL. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, JohnWiley and Sons (1995).

The compounds of this invention may be modified by appending variousfunctionalities via synthetic means delineated herein to enhanceselective biological properties. Such modifications include those whichincrease biological penetration into a given biological system (e.g.,blood, lymphatic system, central nervous system), increase oralavailability, increase solubility to allow administration by injection,alter metabolism and alter rate of excretion.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers. As used herein, the term “pharmaceutically acceptable carrier”means a non-toxic, inert solid, semi-solid or liquid filler, diluent,encapsulating material or formulation auxiliary of any type. Someexamples of materials which can serve as pharmaceutically acceptablecarriers are sugars such as lactose, glucose and sucrose; starches suchas corn starch and potato starch; cellulose and its derivatives such assodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate;powdered tragacanth; malt; gelatin; talc; excipients such as cocoabutter and suppository waxes; oils such as peanut oil, cottonseed oil;safflower oil; sesame oil; olive oil; corn oil and soybean oil; glycols;such a propylene glycol; esters such as ethyl oleate and ethyl laurate;agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator. The pharmaceuticalcompositions of this invention can be administered to humans and otheranimals orally, rectally, parenterally, intracisternally,intravaginally, intraperitoneally, topically (as by powders, ointments,or drops), buccally, or as an oral or nasal spray.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositionswhich can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

Unless otherwise defined, all technical and scientific terms used hereinare accorded the meaning commonly known to one with ordinary skill inthe art. All publications, patents, published patent applications, andother references mentioned herein are hereby incorporated by referencein their entirety.

ABBREVIATIONS

Abbreviations which have been used in the descriptions of the schemesand the examples that follow are:

-   ACN for acetonitrile;-   BOP for benzotriazol-1-yloxy-tris(dimethylamino)phosphonium    hexafluorophosphate;-   Bn for benzyl;-   BTC for bis(trichloromethyl)carbonate; triphosgene;-   BzCl for benzoyl chloride;-   CDI for carbonyldiimidazole;-   DABCO for 1,4-diazabicyclo[2.2.2]octane;-   DAST for diethylaminosulfur trifluoride;-   DABCYL for 6-(N-4′-carboxy-4-(dimethylamino)azobenzene)-aminohexyl-;-   1-O-(2-cyanoethyl)-(N,N-diisopropyl)-phosphoramidite;-   DBU for 1,8-Diazabicycloundec-7-ene;-   DCC for N, N′-dicyclohexylcarbodiimide;-   DCM for dichloromethane;-   DIAD for diisopropyl azodicarboxylate;-   DIBAL-H for diisobutylaluminum hydride;-   DIPEA for diisopropyl ethylamine;-   DMAP for N,N-dimethylaminopyridine;-   DME for ethylene glycol dimethyl ether;-   DMEM for Dulbecco's Modified Eagles Media;-   DMF for N,N-dimethyl formamide;-   DMSO for dimethylsulfoxide;-   DPPA for diphenylphosphoryl azide or diphenyl phosphorylazidate;-   DSC for N, N′-disuccinimidyl carbonate;-   EDCI or EDC for 1-(3-diethylaminopropyl)-3-ethylcarbodiimide    hydrochloride;-   EtOAc for ethyl acetate;-   EtOH for ethyl alcohol;-   HATU for O (7-Azabenzotriazole-1-yl)-N,N,N′,N′-tetramethyluronium    hexafluorophosphate;-   HCl for hydrochloric acid;-   KHMDS is potassium bis(trimethylsilyl) amide;-   PyBrOP for Bromo-tri-pyrolidino-phosphonium hexafluorophosphate;-   Ph for phenyl;-   RT for reverse transcription;-   RT-PCR for reverse transcription-polymerase chain reaction;-   TBME for tert-butyl methyl ether;-   TCDI for 1,1′-thiocarbonyldiimidazole;-   TEA for triethylamine;-   Tf₂O for trifluoromethanesulfonic anhydride;-   TFA for trifluoroacetic acid;-   THE for tetrahydrofuran;-   TLC for thin layer chromatography;-   (TMS)₂NH for hexamethyldisilazane;-   TMSOTf for trimethylsilyl trifluoromethanesulfonate;-   TBS for t-Butyldimethylsilyl;-   TMS for trimethylsilyl;-   TPAP tetrapropylammonium perruthenate;-   TPP or PPh₃ for triphenylphosphine;-   TrCl for trityl chloride;-   Ts for p-CH₃C₆H₄SO₂—-   tBOC or Boc for tert-butyloxy carbonyl; and-   Xantphos for 4,5-Bis-diphenylphosphanyl-9,9-dimethyl-9H-xanthene.    Synthetic Methods

The compounds and processes of the present invention will be betterunderstood in connection with the following synthetic schemes thatillustrate the methods by which the compounds of the invention may beprepared, which are intended as an illustration only and not to limitthe scope of the invention. Various changes and modifications to thedisclosed embodiments will be apparent to those skilled in the art andsuch changes and modifications including, without limitation, thoserelating to the chemical structures, substituents, derivatives, and/ormethods of the invention may be made without departing from the spiritof the invention and the scope of the appended claims.

Scheme 1 illustrates methods for preparing a compound of formula 11,wherein X is OTf or halogen; Y is hydrogen or methyl; n, R₁, m, R₂, v,R₃, ring A and ring B are defined as previously. Deprotection asdescribed in scheme 1 followed by installation of substituted orunsubstituted 4-nitrobenzoyl group provides compound 3. Compound 3 isconverted to vinyl triflate or other vinyl halogen 4, which is coupledwith {circle around (A)}-CO₂Y using a suitable reagent such as a boronicester, boronic acid, organotin reagent, organozinc reagent,organomagnesium reagent or the like, catalyzed by an appropriate Pd, Ni,or Cu catalyst or the like, to afford compounds of formula 5 (5 isconverted to ester 6 if Y is H). Ester 6 is subsequently reduced withreducing agent such as but not limited to, iron, to give compound 7,which is coupled with functionalized acyl chloride 8 to afford compoundsof formula 9. Hydrolysis the ester 9 with a base such as but not limitedto, LiOH or NaOH, gives acid compound 10. Compounds of formula 11 areprepared from the reaction of compound 10 with {circle around (B)}-NH₂.

Scheme 2 illustrates methods to prepare compounds of formula 21, whereinX is OTf or halogen; Y is hydrogen or methyl; n, R₁, m, R₂, v, R₃, ringA and ring B are defined as previously described. Selective methylationof 1 with MeI and a base, such as, but not limited to, t-BuONa, t-BuOKor NaH, gives α-methyl ketone 12. Deprotection as described in Scheme 1followed by installation of substituted or unsubstituted 4-nitrobenzoylgroup provides compound 14. With 14 in hand, compounds of formula 21 areprepared by adapting the similar conditions as described in scheme 1 forcompounds of formula 11.

Scheme 3 illustrates another method to prepare compounds of formula 11,wherein X is OTf or halogen; Y is hydrogen or methyl; n, R₁, m, R₂, v,R₃, ring A and ring B are defined as previously described. Protection of2 with TsCl in the presence of a base, such as but not limited to,pyridine, affords 22 which is converted to vinyl triflate or other vinylhalogen 23. Compound 23 is coupled with {circle around (A)}-CO₂Y using asuitable reagent such as a boronic ester, boronic acid, organotinreagent, organozinc reagent, organomagnesium reagent or the like,catalyzed by an appropriate Pd, Ni, Cu catalyst or the like to afford acompound of formula 24. Deprotection of tosyl group of 24 in acidicconditions such as but not limited to, H₂SO₄, provides 25, which reactswith {circle around (B)}-NH₂ to provide compound 26. Compound 26 iscoupled with functionalized acyl chloride 27 to afford compounds offormula 11.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

Example 1

The synthesis of this compound is illustrated in Scheme 4.

Example 1 Step a

A solution of 1-benzyl-1,2,3,5-tetrahydro-4H-benzo[b]azepin-4-one (3.0g, 11.9 mmol) and Pd/C (2.0 g) in EtOAc/t-BuOH (9:1, 30 mL) was stirredwith hydrogen balloon for 16 h at room temperature. The palladium wasfiltered through Celite and the filtrate was concentrated to afford thedesired product (crude) as yellow oil which was used directly for thenext step. ESI-MS m/z: 162.15 [M+H]⁺.

Example 1 Step b

A solution of the compound from step a (crude), 4-nitrobenzoyl chloride(5.76 g, 31.01 mmol), pyridine (2.45 g, 31 mmol) in CH₂Cl₂ (10 mL) wasstirred for 2 h at room temperature. The resulting solution wasconcentrated by rotary evaporation. The crude product was purified byreverse phase C18 column chromatography (CH₃CN/H₂O) to afford thedesired product (3.4 g) as yellow oil. ESI-MS m/z: 311.05 [M+H]⁺. ¹H NMR(400 MHz, CDCl₃) δ 3.02-2.65 (m, 2H), 3.59 (s, 2H), 4.19 (s, 1H), 4.86(s, 1H), 6.72 (d, J=8.0 Hz, 1H), 7.06 (t, J=8.0 Hz, 1H), 7.23 (t, J=8.0Hz, 1H), 7.32 (d, J=8.0 Hz, 1H), 7.40 (d, J=8.0 Hz, 2H), 8.05 (d, J=8.0Hz, 2H).

Example 1 Step c

A reaction mixture of the compound from step b (3.4 g, 11.0 mmol) andsodium hydride (1.05 g, 43.8 mmol) in DMF (30 mL) was stirred for 30 minat 0° C. Then1,1,1-trifluoro-N-phenyl-N-trifluoromethanesulfonylmethanesulfonamide(4.7 g, 13.1 mmol) was added and the resulting mixture was stirredovernight at room temperature. The residue was purified by reverse flashchromatography (MeCN/H₂O) to give the desired product as a yellow solid(3.6 g, 74%). ESI-MS m/z: 443.10 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 2.92(d, J=16.0 Hz, 1H), 3.09 (t, J=12.0 Hz, 1H), 3.43-3.28 (m, 1H),5.17-5.02 (m, 1H), 6.63 (d, J=8.0 Hz, 1H), 6.75 (s, 1H), 7.02 (t, J=8.0Hz, 1H), 7.23 (t, J=8.0 Hz, 1H), 7.41-7.30 (m, 3H), 8.07 (d, J=8.0 Hz,2H).

Example 1 Step d

A solution of the compound from step c (2.0 g, 4.5 mmol),5-(dihydroxyboranyl)thiophene-2-carboxylic acid (933 mg, 5.4 mmol),Cs₂CO₃ (4.43 g, 13.6 mmol), Pd(PPh₃)₂Cl₂ (317 mg, 0.45 mmol) in DMF (30mL) was heated for 2 h at 80° C. after degassed. The reaction mixturewas used directly for the next step. ESI-MS m/z: 421.10 [M+H]⁺.

Example 1 Step e

The reaction mixture from step d, methyl iodide (2.19 g, 15.46 mmol) inDMF (20 mL) was stirred overnight at room temperature. The residue waspurified by reverse flash chromatography to give the desired product asa yellow solid (1.2 g). ESI-MS m/z: 435.15 [M+H]⁺.

Example 1 Step f

A mixture of the compound from step f (1.20 g, 2.76 mmol), iron (1.54 g,27.62 mmol), NH₄Cl (1.48 g, 27.62 mmol) in EtOH (30 mL) and water (30mL) was heated for 2 h at 80° C. After cooled down, the reaction mixturewas passed through celite. The filtrate was evaporated and the residuewas purified by reverse flash chromatography give the desired product asa yellow solid (950 mg, 85%). ESI-MS m/z: 405.15 [M+H]⁺.

Example 1 Step g

A mixture of 5-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinic acid(405 mg, 1.0 mmol), (1-chloro-2-methylprop-1-en-1-yl)dimethylamine (401mg, 3.0 mmol) in CH₂Cl₂ (30 mL) was stirred for 1 h at room temperature.The solvent was evaporated and the residue was dried in vacuo. Theresidue was dissolved in CH₂Cl₂ (30 mL) and cooled down to 0° C. Afterthat, the compound from step f (263 mg, 1.0 mmol) and pyridine (0.5 mL)were added to the above solution and the mixture was stirred for 1 h atroom temperature. The solvent was removed by rotary evaporation. Theresidue was purified by reverse flash chromatography to give the desiredproduct as a yellow solid (510 mg, 79%). ESI-MS m/z: 649.35 [M+H]⁺.

Example 1 Step h

A mixture of the compound from step g (1.2 g, 1.85 mmol), LiOH (443 mg,18.5 mmol) in methanol (30 mL) and H₂O (30 mL) was stirred for 2 h atroom temperature. The solution was neutralized to pH 7 and solvents wereremoved by rotary evaporation. The residue was purified by reverse flashchromatography to give the desired product as a yellow solid (850 mg,72%). ESI-MS m/z: 635.20 [M+H]⁺.

Example 1 Step i

A mixture of the compound from step h (300 mg, 0.47 mmol) and(1-chloro-2-methylprop-1-en-1-yl)dimethylamine (189 mg, 1.42 mmol) inCH₂Cl₂ (20 mL) was stirred for 30 min at room temperature. The solventwas evaporated and the residue was dried in vacuo. The residue wasdissolved in CH₂Cl₂ (5 mL) and cooled down to 0° C. and thencyclopropanamine (135 mg, 2.36 mmol) and pyridine (0.5 mL) were added.The resulting mixture was stirred for 30 min at room temperature. Afterevaporated the solvent, the residue was purified by reverse flashchromatography to give the desired product as a white solid (230 mg,72%). ESI-MS m/z: 674.40 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 0.61-0.55(m, 2H), 0.76-0.68 (m, 2H), 1.63 (t, J=4.0 Hz, 4H), 2.18 (s, 3H),2.85-2.77 (m, 1H), 3.20-3.00 (m, 4H), 3.46 (t, J=4.0 Hz, 4H), 3.61 (s,4H), 4.95 (s, 1H), 6.72 (d, J=8.0 Hz, 1H), 7.01 (t, J=8.0 Hz, 1H), 7.07(d, J=8.0 Hz, 2H), 7.20 (t, J=8.0 Hz, 1H), 7.26 (s, 1H), 7.38 (d, J=4.0Hz, 1H), 7.56-7.47 (m, 3H), 7.61 (d, J=8.0 Hz, 1H), 7.66 (d, J=4.0 Hz,1H), 8.04 (s, 1H), 8.49 (d, J=4.0 Hz, 1H), 10.38 (s, 1H).

Example 2

Example 2 was prepared using a procedure similar to that used to prepareExample 1, where 2,6-difluoroaniline was used in place ofcyclopropanamine in step i. ESI-MS m/z: 746.40 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 1.63 (t, J=4.0 Hz, 4H), 2.18 (s, 3H), 3.22-3.02 (m, 3H), 3.47(t, J=4.0 Hz, 4H), 3.62 (s, 4H), 4.98 (s, 1H), 6.74 (d, J=8.0 Hz, 1H),7.02 (t, J=8.0 Hz, 1H), 7.09 (d, J=8.0 Hz, 2H), 7.28-7.18 (m, 3H), 7.33(s, 1H), 7.47-7.39 (m, 1H), 7.58-7.48 (m, 4H), 7.64 (d, J=8.0 Hz, 1H),7.97 (d, J=4.0 Hz, 1H), 8.04 (s, 1H), 10.21 (s, 1H), 10.38 (s, 1H).

Example 3

Example 3 was prepared using a procedure similar to that used to prepareExample 1, where cyclobutanamine was used in place of cyclopropanaminein step i. ESI-MS m/z: 688.40 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ1.74-1.58 (m, 6H), 2.14-2.01 (m, 2H), 2.28-2.15 (m, 5H), 3.18-3.01 (m,3H), 3.46 (t, J=4.0 Hz, 4H), 3.61 (s, 4H), 4.45-4.31 (m, 1H), 4.94 (s,1H), 6.72 (d, J=8.0 Hz, 1H), 7.01 (t, J=4.0 Hz, 1H), 7.07 (d, J=8.0 Hz,2H), 7.20 (t, J=4.0 Hz, 1H), 7.26 (s, 1H), 7.40 (d, J=4.0 Hz, 1H),7.57-7.47 (m, 3H), 7.61 (d, J=8.0 Hz, 1H), 7.74 (d, J=4.0 Hz, 1H), 8.05(s, 1H), 8.64 (d, J=8.0 Hz, 1H), 10.38 (s, 1H).

Example 4

Example 4 was prepared using a procedure similar to that used to prepareExample 1, where 3,3-difluorocyclobutan-1-amine was used in place ofcyclopropanamine in step i. ESI-MS m/z: 724.40 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 1.63 (t, J=4.0 Hz, 4H), 2.18 (s, 3H), 2.85-2.68 (m, 2H),3.13-2.90 (m, 5H), 3.46 (t, J=4.0 Hz, 4H), 3.61 (s, 4H), 4.29-4.21 (m,1H), 4.95 (s, 1H), 6.73 (d, J=8.0 Hz, 1H), 7.01 (t, J=8.0 Hz, 1H), 7.07(d, J=8.0 Hz, 2H), 7.20 (t, J=8.0 Hz, 1H), 7.28 (s, 1H), 7.43 (d, J=4.0Hz, 1H), 7.58-7.47 (m, 3H), 7.61 (d, J=8.0 Hz, 1H), 7.73 (d, J=4.0 Hz,1H), 8.29 (s, 1H), 8.87 (t, J=4.0 Hz, 1H), 10.38 (s, 1H).

Example 5

Example 5 was prepared using a procedure similar to that used to prepareExample 1, where bicyclo[1.1.1]pentan-1-amine was used in place ofcyclopropanamine in step i. ESI-MS m/z: 700.40 [M+H]⁺ ¹H NMR (400 MHz,DMSO-d₆) δ 1.66 (t, J=4.0 Hz, 4H), 2.09 (s, 6H), 2.19 (s, 3H), 3.12-3.01(m, 3H), 3.47 (t, J=4.0 Hz, 4H), 3.71 (s, 4H), 4.95 (s, 1H), 6.72 (d,J=8.0 Hz, 1H), 7.00 (t, J=8.0 Hz, 1H), 7.09 (d, J=8.0 Hz, 2H), 7.19 (t,J=8.0 Hz, 1H), 7.25 (s, 1H), 7.38 (d, J=4.0 Hz, 1H), 7.53 (d, J=8.0 Hz,2H), 7.61 (d, J=8.0 Hz, 2H), 7.72-7.65 (m, 2H), 8.02 (s, 1H), 8.99 (s,1H), 10.48 (s, 1H).

Example 6

Example 6 was prepared using a procedure similar to that used to prepareExample 1, where 2-fluoro-6-methylaniline was used in place ofcyclopropanamine in step i. ESI-MS m/z: 742.40 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 1.63 (t, J=4.0 Hz, 4H), 2.18 (s, 3H), 2.25 (s, 3H), 3.19-3.05(m, 3H), 3.47 (t, J=4.0 Hz, 4H), 3.62 (s, 4H), 4.97 (s, 1H), 6.74 (d,J=8.0 Hz, 1H), 7.02 (t, J=8.0 Hz, 1H), 7.09 (t, J=8.0 Hz, 2H), 7.16 (d,J=8.0 Hz, 2H), 7.20 (t, J=8.0 Hz, 1H), 7.34-7.25 (m, 2H), 7.49 (d, J=4.0Hz, 2H), 7.54 (d, J=8.0 Hz, 2H), 7.63 (d, J=8.0 Hz, 1H), 7.95 (s, 1H),8.04 (s, 1H), 9.96 (s, 1H), 10.38 (s, 1H).

Example 7

Example 7 was prepared using a procedure similar to that used to prepareExample 2, where 6-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinicacid was used in place of5-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinic acid in step g.ESI-MS m/z: 746.45 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 1.64 (t, J=4.0Hz, 4H), 2.36 (s, 3H), 3.22-3.02 (m, 3H), 3.48 (t, J=4.0 Hz, 4H), 3.68(s, 4H), 4.97 (s, 1H), 6.60 (d, J=8.0 Hz, 1H), 6.74 (d, J=8.0 Hz, 1H),7.02 (t, J=8.0 Hz, 1H), 7.08 (t, J=8.0 Hz, 2H), 7.28-7.17 (m, 3H), 7.33(s, 1H), 7.47-7.39 (m, 1H), 7.51 (d, J=4.0 Hz, 1H), 7.54 (d, J=8.0 Hz,2H), 7.68-7.57 (m, 2H), 7.97 (d, J=4.0 Hz, 1H), 10.22 (s, 1H), 10.35 (s,1H).

Example 8

Example 8 was prepared using a procedure similar to that used to prepareExample 6, where 6-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinicacid was used in place of5-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinic acid in step g.ESI-MS m/z: 742.50 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 1.63 (t, J=4.0Hz, 4H), 2.25 (s, 3H), 2.33 (s, 3H), 3.19-3.05 (m, 3H), 3.47 (t, J=4.0Hz, 4H), 3.64 (s, 4H), 4.97 (s, 1H), 6.57 (d, J=8.0 Hz, 1H), 6.74 (d,J=8.0 Hz, 1H), 7.02 (t, J=8.0 Hz, 1H), 7.08 (t, J=8.0 Hz, 2H), 7.16 (d,J=8.0 Hz, 2H), 7.21 (t, J=8.0 Hz, 1H), 7.34-7.25 (m, 2H), 7.49 (d, J=4.0Hz, 1H), 7.54 (d, J=8.0 Hz, 3H), 7.63 (d, J=8.0 Hz, 1H), 7.95 (s, 1H),9.96 (s, 1H), 10.38 (s, 1H).

Example 9

Example 9 was prepared using a procedure similar to that used to prepareExample 4, where 6-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinicacid was used in place of5-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinic acid in step g.ESI-MS m/z: 724.45 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 1.63 (t, J=4.0Hz, 4H), 2.33 (s, 3H), 2.85-2.60 (m, 2H), 3.20-2.90 (m, 5H), 3.47 (t,J=4.0 Hz, 4H), 3.64 (s, 4H), 4.35-4.15 (m, 1H), 4.97 (s, 1H), 6.57 (d,J=8.0 Hz, 1H), 6.72 (d, J=8.0 Hz, 1H), 7.15-7.01 (m, 3H), 7.22 (t, J=8.0Hz, 1H), 7.26 (s, 1H), 7.40 (d, J=4.0 Hz, 1H), 7.56-7.50 (m, 3H), 7.60(d, J=8.0 Hz, 1H), 7.74 (d, J=4.0 Hz, 1H), 8.84 (d, J=8.0 Hz, 1H), 10.26(s, 1H).

Example 10

Example 10 was prepared using a procedure similar to that used toprepare Example 3, where6-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinic acid was used inplace of 5-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinic acid instep g. ESI-MS m/z: 688.50 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 1.74-1.58(m, 6H), 2.14-2.02 (m, 2H), 2.27-2.16 (m, 2H), 2.33 (s, 3H), 3.14-2.98(m, 3H), 3.47 (t, J=4.0 Hz, 4H), 3.64 (s, 4H), 4.45-4.32 (m, 1H), 4.95(s, 1H), 6.57 (d, J=8.0 Hz, 1H), 6.72 (d, J=8.0 Hz, 1H), 7.01 (t, J=8.0Hz, 1H), 7.07 (t, J=8.0 Hz, 2H), 7.20 (t, J=8.0 Hz, 1H), 7.26 (s, 1H),7.40 (d, J=4.0 Hz, 1H), 7.56-7.50 (m, 3H), 7.60 (d, J=8.0 Hz, 1H), 7.74(d, J=4.0 Hz, 1H), 8.63 (d, J=8.0 Hz, 1H), 10.29 (s, 1H).

Example 11

Example 11 was prepared using a procedure similar to that used toprepare Example 3, where6-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinic acid was used inplace of 5-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinic acid instep g. ESI-MS m/z: 700.45 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 1.63 (t,J=4.0 Hz, 4H), 2.09 (s, 6H), 2.33 (s, 3H), 2.47 (s, 1H), 3.18-2.98 (m,3H), 3.47 (t, J=4.0 Hz, 4H), 3.64 (s, 4H), 4.94 (s, 1H), 6.57 (d, J=8.0Hz, 1H), 6.72 (d, J=8.0 Hz, 1H), 7.01 (t, J=8.0 Hz, 1H), 7.07 (d, J=8.0Hz, 2H), 7.19 (t, J=8.0 Hz, 1H), 7.25 (s, 1H), 7.39 (d, J=4.0 Hz, 1H),7.57-7.50 (m, 3H), 7.61 (d, J=8.0 Hz, 1H), 7.69 (d, J=4.0 Hz, 1H), 8.99(s, 1H), 10.30 (s, 1H).

Example 12

Example 12 was prepared using a procedure similar to that used toprepare Example 1, where methyl5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiophene-3-carboxylatewas used in place of 5-(dihydroxyboranyl)thiophene-2-carboxylic acid instep b. ES-MS m/z 674.35 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 0.51-0.60(m, 2H), 0.71 (dt, J=6.9, 3.3 Hz, 2H), 1.66 (t, J=5.2 Hz, 4H), 2.19 (s,3H), 2.81 (m, 1H), 3.05 (s, 3H), 3.47 (t, J=5.1 Hz, 4H), 3.70 (s, 4H),4.95 (s, 1H), 6.73 (d, J=7.9 Hz, 1H), 7.00 (t, J=7.6 Hz, 1H), 7.08 (d,J=8.3 Hz, 2H), 7.15 (s, 1H), 7.19 (td, J=7.6, 1.3 Hz, 1H), 7.53 (d,J=8.3 Hz, 2H), 7.56-7.62 (m, 1H), 7.66 (s, 1H), 7.73 (d, J=1.4 Hz, 1H),8.03 (dd, J=4.7, 1.7 Hz, 2H), 8.26 (d, J=4.1 Hz, 1H), 10.47 (s, 1H).

Example 13

Example 13 Step a

To a round-bottom flask (100 mL) were added1-benzyl-3,5-dihydro-2H-1-benzazepin-4-one (1.50 g, 5.97 mmol), t-BuONa(0.57 g, 5.97 mmol), CH₃I (0.85 g, 5.97 mmol) and THE (40 mL) at roomtemperature. The resulting mixture was stirred for 20 min at 70° C. andthen diluted with brine (50 mL). After extracted with EtOAc (3×100 mL),the combined organic layers were dried, evaporated and purified byreverse flash chromatography to afford the crude product (1.1 g) asyellow oil. ESI-MS m/z: 266.25 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 1.34(d, J=7.0 Hz, 3H), 2.29 (dt, J=12.5, 6.0 Hz, 1H), 2.58 (dt, J=12.9, 7.6Hz, 1H), 3.10 (dd, J=7.5, 6.0 Hz, 2H), 3.84 (q, J=7.0 Hz, 1H), 4.21-4.36(m, 2H), 7.03 (td, J=7.3, 1.4 Hz, 1H), 7.13-7.27 (m, 4H), 7.28-7.38 (m,4H).

Example 13 Step b

To a round-bottom flask (100 mL) were added the compound from step a(1.10 g, 4.14 mmol), EtOAc (40 mL) and t-BuOH (5 mL) at roomtemperature. Then Pd/C (2.21 g) was added under nitrogen atmosphere atroom temperature and the resulting mixture was stirred for 2 h at roomtemperature under H₂ atmosphere. The reaction mixture was passed throughCelite and the filter cake was washed with EtOAc (3×10 mL). The filtratewas concentrated under reduced pressure to give the crude product (800mg) as yellow oil. ESI-MS m/z: 176.10 [M+H]⁺.

Example 13 Step c

To a round-bottom flask (100 mL) were added the compound from step b(800 mg, 4.56 mmol), 4-nitro-(1.27 g, 6.84 mmol), pyridine (722 mg, 9.13mmol) and CH₂Cl₂ (35 mL) at 0° C. The resulting mixture was stirred for30 min at room temperature. Then the mixture was concentrated underreduced pressure and the residue was purified by reverse flash andPrep-TLC (CH₂Cl₂/MeOH 15:1) to afford the desired product (900 mg) as ayellow solid. ESI-MS m/z: 325.15 [M+H]⁺.

Example 13 Step d

To a round-bottom flask (100 mL) were added the compound from step c(1.14 g, 3.51 mmol) and DMF (40 mL) at room temperature. After thesolution was cooled down to 0° C., t-BuONa (1.01 g, 10.54 mmol) and1,1,1-trifluoro-N-phenyl-N-trifluoromethanesulfonylmethane-sulfonamide(2.51 g, 7.02 mmol) were added. The resulting mixture was slowly warmedto room temperature and stirred further for 30 min. The reaction mixturewas quenched with water and extracted with EtOAc (5×200 mL). Afterseparation, the combined organic layers were dried, filtered andconcentrated under reduced pressure. The residue was purified by reverseflash chromatography to afford the desired product (1.0 g) as a brownyellow solid. ESI-MS m/z: 457.05[M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 2.30(d, J=1.6 Hz, 3H), 2.57-2.82 (m, 2H), 3.71 (ddd, J=13.0, 6.3, 2.8 Hz,1H), 4.87 (td, J=12.6, 6.1 Hz, 1H), 7.04 (dd, J=7.9, 1.4 Hz, 1H), 7.13(td, J=7.6, 1.5 Hz, 1H), 7.23-7.41 (m, 3H), 7.59 (dd, J=7.9, 1.5 Hz,1H), 8.00-8.13 (m, 2H).

Example 13 Step e

To a round-bottom flask (100 mL) were added the compound from step d(500 mg, 1.09 mmol), 5-(dihydroxyboranyl)thiophene-2-carboxylic acid(282 mg, 1.64 mmol), Pd(dppf)Cl₂ (80 mg, 0.11 mmol), Cs₂CO₃ (714 mg,2.19 mmol) and DMF (40 mL) at room temperature. After degassed, themixture was heated for 2 h at 100° C. The resulting mixture was used innext step without purification. ESI-MS m/z: 435.10[M+H]⁺.

Example 13 Step f

The mixture from step e was cooled down to room temperature, and thenCH₃I (1 mL) was added. The resulting mixture was stirred for 1 h at roomtemperature. The solid was filtered out through celite. The filter cakewas washed with EtOAc (3×30 mL). The filtrate was concentrated underreduced pressure. The residue was purified by reverse flashchromatography to afford the crude product (280 mg) as a dark greensolid. ESI-MS m/z: 449.10 [M+H]⁺.

Example 13 Step g

To a round-bottom flask (100 mL) were added the compound from step f(260 mg, 0.58 mmol), Fe (323 mg, 5.79 mmol), NH₄Cl (310 mg, 5.79 mmol),H₂O (3 mL) and EtOH (25 mL) at room temperature. The resulting mixturewas heated for 30 min at 80° C. After cooled down, the mixture wasconcentrated under vacuum. The stick residue was washed with CH₂Cl₂(4×50 mL). The combined CH₂Cl₂ was passed through celite and thefiltrate was concentrated under reduced pressure. The obtained residuewas purified by prep-TLC (CH₂Cl₂/MeOH 15:1) to afford the desiredproduct (220 mg) as a yellow solid. ESI-MS m/z: 419.10 [M+H]⁺.

Example 13 Step h

A mixture of 5-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinic acid(179 mg, 0.68 mmol) and (1-chloro-2-methylprop-1-en-1-yl)dimethylamine(210 mg, 1.57 mmol) in CH₂Cl₂ (10 mL) was stirred for 1 h at roomtemperature under nitrogen atmosphere. The resulting mixture wasconcentrated and the residue was dried in vacuo. The residue wasdissolved in CH₂Cl₂ (5 mL) and cooled down to 0° C. To the solution wereadded the compound from step g (220 mg, 0.52 mmol), CH₂Cl₂ (10 mL) andpyridine (1 mL). The resulting mixture was stirred for 2 h at roomtemperature and then concentrated. The obtained residue was purified byprep-TLC (CH₂Cl₂/MeOH 15:1) to get the desired product (300 mg) as ayellow solid. ESI-MS m/z: 663.20 [M+H]⁺.

Example 13 Step i

To a 100 mL round-bottom flask were added the compound from step h (300mg, 0.45 mmol), MeOH (30 mL) and a solution of LiOH (108 mg, 4.52 mmol)in H₂O (5 mL). The mixture was stirred overnight at room temperature andthen acidified to pH 5 with HCl (2M aq.). The mixture was concentratedunder reduced pressure and the residue was purified by reverse flashchromatography to afford the desired product (270 mg) as a yellow solid.ESI-MS m/z: 649.35 [M+H]⁺.

Example 13 Step j

To a sealed tube (8 mL) were added the compound from step i (60 mg, 0.09mmol), (1-chloro-2-methylprop-1-en-1-yl)dimethylamine (37 mg, 0.27 mmol)and CH₂Cl₂ (2 mL) at room temperature. The resulting mixture was stirredfor 1 h at room temperature. The resulting mixture was concentrated andthe residue was dried in vacuo. The residue was dissolved in CH₂Cl₂ (2mL) and cooled down to 0° C. To the mixture were added cyclopropanamine(31 mg, 0.55 mmol) and pyridine (0.5 mL) which was stirred for 1 h atroom temperature. After evaporated the solvent, the residue was purifieddirectly by prep-TLC (CH₂Cl₂/MeOH 15:1) and prep-HPLC to afford thedesired product as a white solid. ESI-MS m/z: 688.50 [M+H]⁺. ¹H NMR (400MHz, DMSO-d₆) δ 0.55-0.62 (m, 2H), 0.71 (dt, J=6.9, 3.4 Hz, 2H), 1.63(t, J=5.2 Hz, 4H), 2.17 (s, 3H), 2.40 (s, 4H), 2.65 (d, J=17.8 Hz, 1H),2.81 (td, J=7.2, 3.7 Hz, 1H), 3.47 (t, J=5.2 Hz, 4H), 3.61 (s, 4H), 3.76(s, 1H), 4.56 (s, 1H), 6.87 (s, 1H), 7.08 (s, 3H), 7.31 (d, J=8.9 Hz,2H), 7.40-7.58 (m, 4H), 7.72 (d, J=4.0 Hz, 1H), 8.04 (d, J=2.2 Hz, 1H),8.38-8.54 (m, 1H), 10.35 (s, 1H).

Example 14

Example 14 was prepared using a procedure similar to that used toprepare Example 13, where 2-fluoro-6-methylaniline was used in place ofcyclopropanamine. ESI-MS m/z: 756.50 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ1.63 (t, J=5.1 Hz, 4H), 2.17 (s, 3H), 2.26 (s, 3H), 2.43 (s, 4H), 2.67(s, 1H), 3.47 (d, J=5.3 Hz, 4H), 3.61 (s, 4H), 3.77 (d, J=13.6 Hz, 1H),4.60 (s, 1H), 6.89 (s, 1H), 7.13 (dd, J=19.5, 10.4 Hz, 5H), 7.24-7.42(m, 3H), 7.43-7.59 (m, 4H), 8.02 (dd, J=18.2, 3.0 Hz, 2H), 9.98 (s, 1H),10.35 (s, 1H).

Example 15

Example 15 Step a

A solution of the compound from Example 1 step a (1.35 g, 8.35 mmol) andTsCl (3.5 g, 18.61 mmol) in pyridine (10 mL) was stirred for 2 h at roomtemperature. The resulting solution was extracted with EtOAc (3×50 mL).The combined organic layer was washed with brine, dried, filtered andconcentrated. The crude product was purified by reverse phase C18 columnchromatography (CH₃CN/H₂O) to afford the desired product (1.4 g) asyellow oil. ESI-MS m/z: 316.20 [M+H]⁺.

Example 15 Step b

A solution of the compound of step a (1.4 g, 4.44 mmol) in THE (50 mL)was cooled downed to −78° C. and LiHMDS (5.3 mL, 5.33 mmol) was added.The resulting mixture was stirred for 1 h at −78° C. After that,1,1,1-trifluoro-N-phenyl-N-trifluoromethanesulfonylmethanesulfonamide(1.5 g, 4.18 mmol) in THE (20 mL) was gradually added into the solutionand stirred for 1 h. The resulting solution was diluted with saturatedNH₄Cl solution and extracted with EtOAc (3×50 mL). The combined organiclayer was washed with brine, dried, filtered and concentrated. The crudeproduct was purified by reverse phase C18 column chromatography(CH₃CN/H₂O) to afford the desired product (980 mg, 49%) as yellow oil.ESI-MS m/z: 448.15 [M+H]⁺.

Example 15 Step c

A solution of the compound from step b (150 mg, 0.33 mmol) and4-(methoxycarbonyl)phenylboronic acid (120 mg, 0.67 mmol), Pd(PPh₃)₂Cl₂(47 mg, 0.07 mmol) and Cs₂CO₃ (328 mg, 1.00 mmol) in DMF (3 mL) with aninert atmosphere of nitrogen was heated for 2 h at 80° C. The resultingsolution was extracted with EtOAc (3×20 mL), and the combined organiclayer was washed with brine, dried, filtered and concentrated to affordthe desired product (crude) as a yellow solid. ESI-MS m/z: 434.25[M+H]⁺.

Example 15 Step d

A solution of the compound from step c (crude) in H₂SO₄ (90%) (4.00 mL)was heated for 16 h at 70° C. The pH of the resulting solution wasadjusted to ˜6 with NaHCO₃ (aq) and then extracted with EtOAc (3×20 mL).The combined organic layer was washed with brine, dried, filtered andconcentrated. The crude product was purified by reverse phase C18 columnchromatography (CH₃CN/H₂O) to afford the desired product (50 mg, 56%) asa yellow solid. ESI-MS m/z: 266.20 [M+H]⁺.

Example 15 Step e

A solution of the compound from step d (50 mg, 0.19 mmol),cyclopropanamine (54 mg, 0.94 mmol), DIPEA (73 mg, 0.56 mmol) and HATU(107 mg, 0.28 mmol) in DMF (2 mL) was stirred for 2 h at roomtemperature. The solution was purified by reverse phase C18 columnchromatography (CH₃CN/H₂O) to give the desired compound as a white solid(40 mg, 69%). ESI-MS m/z: 305.25 [M+H]⁺.

Example 15 Step f

A solution of4-(5-methyl-2-(7-oxa-2-azaspiro[3.5]nonan-2-yl)nicotinamido)benzoic acid(100 mg, 0.26 mmol) and (1-chloro-2-methylprop-1-en-1-yl)dimethylamine(105 mg, 0.79 mmol) in CH₂Cl₂ (10 mL) was stirred for 2 h at roomtemperature. Then the resulting mixture was concentrated under vacuumand the residue was dried in vacuo. The residue was dissolved in CH₂Cl₂(5 mL) and cooled down to 0° C. After that, the compound from step e (40mg, 0.13 mmol) and pyridine (31 mg, 0.39 mmol) in CH₂Cl₂ (10 mL) wereadded. The resulting solution was stirred for 1 h at 0° C. and thenconcentrated. The crude product was purified by Prep-HPLC (CH₃CN/H₂O) toafford the desired product (25.3 mg, 23%) as a yellow solid. ESI-MS m/z:668.50 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 0.61 (s, 2H), 0.68-0.74 (m,2H), 1.65 (s, 4H), 2.19 (s, 3H), 2.68 (s, 1H), 3.05 (s, 2H), 3.47 (s,6H), 3.65 (s, 4H), 4.95 (s, 1H), 6.74 (d, J=8.6 Hz, 1H), 7.00 (t, J=8.0Hz, 1H), 7.11 (d, J=6.7 Hz, 3H), 7.20 (t, J=7.5 Hz, 1H), 7.54 (d, J=8.3Hz, 2H), 7.61 (d, J=7.3 Hz, 1H), 7.75 (d, J=8.2 Hz, 2H), 7.88 (d, J=8.2Hz, 2H), 8.04 (s, 1H), 8.47 (d, J=4.4 Hz, 1H), 10.41 (s, 1H).

Example 16

Example 16 was prepared using a procedure similar to that used toprepare Example 15, where 2-fluoro-6-methylaniline was used in place ofcyclopropanamine in step e. ESI-MS m/z: 736.40 [M+H]⁺. ¹H NMR (400 MHz,DMSO-d₆) δ 1.64 (t, J=5.2 Hz, 4H), 2.18 (s, 3H), 2.26 (s, 3H), 3.09 (s,3H), 3.47 (t, J=5.1 Hz, 4H), 3.62 (s, 4H), 5.00 (s, 1H), 6.75 (d, J=7.9Hz, 1H), 7.02 (t, J=7.7 Hz, 1H), 7.09-7.32 (m, 7H), 7.47-7.59 (m, 3H),7.60-7.66 (m, 1H), 7.84 (d, J=8.2 Hz, 2H), 8.02-8.09 (m, 3H), 9.94 (s,1H), 10.39 (s, 1H).

Example 17

Example 17 was prepared using a procedure similar to that used toprepare Example 6, where 2-fluoro-6-methylaniline was used in place of5-(dihydroxyboranyl)thiophene-2-carboxylic acid in step d. ESI-MS m/z:754.35 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 1.67 (t, J=5.2 Hz, 4H), 2.20(s, 3H), 2.26 (s, 3H), 2.95 (d, J=71.7 Hz, 3H), 3.48 (t, J=5.3 Hz, 4H),3.71 (s, 4H), 4.95 (s, 1H), 6.78 (d, J=8.0 Hz, 1H), 6.86 (s, 1H), 7.04(t, J=7.6 Hz, 1H), 7.16 (t, J=8.9 Hz, 4H), 7.21 (t, J=7.5 Hz, 1H),7.25-7.34 (m, 1H), 7.55 (t, J=8.5 Hz, 3H), 7.67 (s, 1H), 7.74 (t, J=7.9Hz, 1H), 7.90 (dd, J=16.6, 9.8 Hz, 2H), 8.04 (d, J=2.2 Hz, 1H), 10.05(s, 1H), 10.50 (s, 1H).

Example 18

Example 18 was prepared using a procedure similar to that used toprepare Example 6, where 2-chloro-6-methylaniline was used in place of5-(dihydroxyboranyl)thiophene-2-carboxylic acid in step d. ESI-MS m/z:770.50 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 1.64 (t, J=4.0 Hz, 4H), 2.18(s, 3H), 2.26 (s, 3H), 3.19-2.73 (m, 3H), 3.48 (t, J=4.0 Hz, 4H), 3.63(s, 4H), 4.96 (s, 1H), 6.62 (s, 1H), 6.78 (d, J=8.0 Hz, 1H), 7.03 (t,J=8.0 Hz, 1H), 7.22-7.11 (m, 5H), 7.32-7.24 (m, 1H), 7.52-7.46 (m, 2H),7.56 (d, J=8.0 Hz, 2H), 7.67 (d, J=8.0 Hz, 1H), 8.08-7.98 (m, 2H), 8.14(s, 1H), 10.08 (s, 1H), 10.39 (s, 1H).

Example 19

Example 19 was prepared using a procedure similar to that used toprepare Example 6, where 2-methyl-6-methylaniline was used in place of5-(dihydroxyboranyl)thiophene-2-carboxylic acid in step d. ESI-MS m/z:750.60 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆) δ 1.64 (t, J=4.0 Hz, 4H), 2.18(s, 3H), 2.25 (s, 3H), 2.40 (s, 3H), 3.20-2.72 (m, 3H), 3.48 (t, J=4.0Hz, 4H), 3.63 (s, 4H), 4.93 (s, 1H), 6.51 (s, 1H), 6.77 (d, J=8.0 Hz,1H), 7.01 (t, J=8.0 Hz, 1H), 7.21-7.10 (m, 5H), 7.42 (d, J=8.0 Hz, 1H),7.47 (d, J=4.0 Hz, 1H), 7.51 (s, 1H), 7.57 (d, J=8.0 Hz, 2H), 7.86 (d,J=8.0 Hz, 1H), 7.91 (s, 1H), 8.05 (s, 1H), 9.88 (s, 1H), 10.40 (s, 1H).

Assays

Introduction

RSV is a single stranded negative sense RNA virus that causesrespiratory tract infections which can be dangerous to infants, theelderly, and immunosuppressed individuals. Currently there is novaccine, and therapeutic options are both costly and of limitedeffectiveness. These approved treatments are Ribavirin, andPalivizumab/Synagis (a monoclonal antibody). RSV has two genotypes, Aand B, which differ primarily in the structure of the virus' surface “G”attachment protein. Our current primary screen focusses on RSV-A anduses an in vitro cytoprotection assay where compounds are added in2-fold dilutions to cells which are then subjected to fully replicativeviral particles. Cell viability is measured several days later alongwith separate measurements of compound cytotoxicity. This report focuseson the results of our most recent screening of compounds.

Methods

HEp-2 cells, (originally derived from tumors grown inirradiated-cortisonised weanling rats that had been injected withepidermoid carcinoma tissue from a 56 year old male's larynx, but laterfound to be indistinguishable from HeLa cells by PCR DNA analysis), wereused for the culturing of genotype A, “Long” strain RSV. Flasks wereinoculated with RSV and viral stocks were collected once cytopathiceffect (CPE) was greater than 90%. Viral stocks in 25% sucrose mediawere snap frozen using liquid nitrogen to increase viral stability.Viral stock titers were quantified by tissue culture infectious dose 50%(TCID₅₀) using 8,000 cells per well and 3-fold viral dilutions across a96-well plate, cultured for 4 days.

The control compound currently used in the RSV assay is RSV-604, a ˜2.4μM EC₅₀ nucleocapsid inhibitor previously developed by Novartis.Following extensive parameter testing, the final assay is run asfollows: HEp-2 cells are seeded into the inner 60 wells of a 96-wellplate at 8,000 cells per well in a volume of 50 μL using Growth Media(DMEM without phenol red, 1% L-Glut, 1% Penn/Strep, 1% nonessentialamino acids, 10% FBS). 2-Fold serial dilutions of control and testcompounds are added to the wells in duplicate in a total volume of 25μL. Viral stock is then added to the wells in a volume of 25 μL,bringing the total volume of each well to 100 μL. Each 96-well plate hasa control column of 6 wells with cells and virus but no compound(negative control, max CPE), a column with cells but no compound orvirus (positive control, minimum CPE), and a column with no cells orvirus or compound (background plate/reagent control). The control wellswith cells but no virus are given an additional 25 μL of growth mediacontaining an equal quantity of sucrose as those wells receiving theviral stock in order to keep consistent in media and volume conditions.The outer wells of the plate are filled with 125 μL of growth media toact as a thermal and evaporative moat around the test wells. Following a4-day incubation period, the plates are read using ATPlite (50 uL addedper well), which quantifies the amount of ATP (a measure of cell health)present in each well. Assay plates are read using the Envisionluminometer. In parallel, cytotoxicity is examined on an additional96-well plate treated in an identical manner, but substituting the 25 μLof viral stock for 25 μL of growth media. These data are used tocalculate the EC₅₀ of each compound. EC₅₀ ranges are as follows: A<0.01μM; B 0.01-0.05 μM; C>0.05 μM.

TABLE 1 Summary of Activities Human RSV-A Human RSV-A (“Long” strain)(“Long” strain) Example EC₅₀ Example EC₅₀ 1 A 2 A 3 A 4 A 5 B 6 B 7 B 8B 9 B 10 A 11 B 12 B 13 C 14 C 15 A 16 A

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

What is claimed is:
 1. A compound represented by Formula (I):

or a pharmaceutically acceptable salt thereof, wherein: A is selectedfrom the group consisting of: 1) optionally substituted aryl; 2)optionally substituted heteroaryl; 3) optionally substituted —C₃-C₁₂cycloalkyl; 4) optionally substituted —C₃-C₁₂ cycloalkenyl; and 5)optionally substituted 3- to 12-membered heterocycloalkyl; B is selectedfrom the group consisting of: 1) optionally substituted aryl; 2)optionally substituted heteroaryl; 3) optionally substituted —C₃-C₁₂cycloalkyl; 4) optionally substituted —C₃-C₁₂ cycloalkenyl; and 5)optionally substituted 3- to 12-membered heterocycloalkyl; L is absent,—CONH—, —NHCO—, —NHCO₂—, or —NHS(O)₂—; Each R₁, R₂, and R₃ isindependently selected from the group consisting of: 1) halogen; 2)cyano; 3) nitro; 4) optionally substituted —C₁-C₈ alkoxy; 5) optionallysubstituted —C₁-C₈ alkyl; 6) optionally substituted —C₂-C₈ alkenyl; 7)optionally substituted —C₂-C₈ alkynyl; 8) optionally substituted —C₃-C₆cycloalkyl; 9) optionally substituted —C₃-C₆ cycloalkenyl; and 10)optionally substituted 3- to 6-membered heterocycloalkyl; R₄ is selectedfrom the group consisting of: 1) hydrogen; 2) halogen; 3) cyano; 4)nitro; 5) optionally substituted —C₁-C₈ alkoxy; 6) optionallysubstituted —C₁-C₈ alkyl; 7) optionally substituted —C₂-C₈ alkenyl; 8)optionally substituted —C₂-C₈ alkynyl; 9) optionally substituted —C₃-C₈cycloalkyl; 10) optionally substituted —C₃-C₈ cycloalkenyl; and 11)optionally substituted 3- to 8-membered heterocycloalkyl; n is 0, 1, 2,3 or 4; m is 0, 1, 2, 3 or 4; and v is 0, 1, 2, or
 3. 2. The compound ofclaim 1, wherein L is absent or —CONH—.
 3. The compound of claim 1,wherein A is selected from one of the following groups by removal of twohydrogen atoms, wherein each of the groups is optionally substitutedwhen possible:


4. The compound of claim 1, wherein A is selected from the followinggroups, wherein each group is optionally substituted:


5. The compound of claim 1, wherein B is selected from the followinggroups by removal of a hydrogen atom, wherein each group is optionallysubstituted when possible:


6. The compound of claim 1, represented by one of Formulas (IIa), (IIb,and (IIc), or a pharmaceutically acceptable salt thereof:

wherein A, B, R₁, R₂, R₃, R₄, n, m, and v are as defined in claim
 1. 7.The compound of claim 1, represented by one of Formulas (IVa), (IVb),and (IVc), or a pharmaceutically acceptable salt thereof:

wherein A, B, R₁, R₃, and R₄ are as defined in claim
 1. 8. The compoundof claim 1, represented by one of Formulas (Va), (Vb), and (Vc), or apharmaceutically acceptable salt thereof:

wherein A, B, R₃, and R₄ are as defined in claim
 1. 9. The compound ofclaim 1, represented by one of Formulas (VIa), (VIb), and (VIc), or apharmaceutically acceptable salt thereof:

wherein B, R₁, R₃, and R₄ are as defined in claim
 1. 10. The compound ofclaim 1, represented by one of Formulas (VIIa), (VIIb), and (VIIc), or apharmaceutically acceptable salt thereof:

wherein B, R₃, and R₄ are as defined in claim
 1. 11. The compound ofclaim 1, selected from the compounds set forth below or apharmaceutically acceptable salt thereof: Compound Structure  1

 2

 3

 4

 5

 6

 7

 8

 9

10

11

12

13

14

15

16

17

18

19


12. A pharmaceutical composition comprising a compound according toclaim 1 and a pharmaceutically acceptable carrier, diluent or excipient.13. A method of treating or preventing an RSV infection in a subject inneed thereof, comprising administering to the subject a therapeuticallyeffective amount of a compound of claim
 1. 14. The method of claim 13,further comprising the step of administering to the subject anadditional anti-RSV agent.
 15. The method of claim 13, furthercomprising administering to the subject a steroid anti-inflammatorycompound.
 16. A method of treating RSV and influenza in a subject inneed thereof, comprising administering to the subject a therapeuticallyeffective amount of a compound of claim 1 and a therapeuticallyeffective amount of an anti-influenza agent.
 17. The method of claim 14,wherein the compound and the additional anti-RSV agent areco-formulated.
 18. The method of claim 14, wherein the compound and theadditional anti-RSV agent are co-administered.
 19. The method of claim14, wherein the additional anti-RSV agent is administered at a lowerdose and/or frequency compared to the dose and/or frequency with whichit is administered as a monotherapy.